The_Determination_of_the_Isoelectric_Point_of_the_Protein_Bovine_Serum_Albumin[1] - Analytical Che

The_Determination_of_the_Isoelectric_Point_of_the_Protein_Bovine_Serum_Albumin[1] - Analytical Che

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The Determination of the Isoelectric Point of the Protein Bovine Serum Albumin Flora Vo Lab Partners: Taylor Scott, Tessa Lindsay, and Luke Machen 22 February 2007 TA: BJ Privett Section 410: Thursday 1:00 – 4:00 PM Pledge: I pledge that no unauthorized assistance has been given or received in the completion of the work presented in this report.
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Introduction Using electrophoresis, the objective of this lab is to determine the isoelectric point of bovine serum albumin (BSA). Electrophoresis employs an electric field and a buffer solution to separate molecules based on charge and size. This experiment will use slab electrophoresis separation, which utilizes a thin strip of electrophoresis paper made from cellulose polyacetate. The paper is porous and semi-solid to allow the substance of interest to migrate. The substance, shaped as a spot or a line, is applied in the middle of the strip; this is called the sample origin. To perform electrophoresis, this piece of paper will have its ends immersed in the buffer solution, with one end in the cathode reservoir and one end in the anode reservoir. The wetted strip connects the two reservoirs, like a salt bridge. When a voltage is provided, an electric field is created, utilizing the aqueous solution containing negative and positive charges. The molecule of interest will then migrate toward the cathode or anode. The formula for the electric field is given below: with V = voltage and d = distance between the two electrodes E = V/d The direction of migration depends on the charge of the molecule. A molecule with a positive charge will migrate toward the cathode, which is negative. A molecule with a negative charge will migrate toward the anode, which is positive. An overall neutral molecule will remain at the sample origin. Simply, opposites attract. The rate of migration depends on the electric field (E) and electrophilic mobility ( μ ep ), which consists of charge (q) and the frictional coefficient (f). The frictional coefficient is a complex variable, whose value depends on hydrodynamic radius, shape, and viscosity. U ep = μ ep E => uep = (q/f)E From this equation, the higher charge the molecule has, the further it will migrate, and vice
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versa. The lower the frictional forces, which include size and weight, the further it will travel also. If the molecule is large and was a high molecular weight, it will not travel far. In conclusion, the charge and size influence how far and in which direction the substance will migrate. Amino acids have at least two hydrogen atoms to denote as the pH increases, and
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The_Determination_of_the_Isoelectric_Point_of_the_Protein_Bovine_Serum_Albumin[1] - Analytical Che

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